Cargando…

Enzymatic Phosphorylation of Oxidized Tyrosine Residues

[Image: see text] Post-translational modifications (PTMs) alter the function and fate of proteins and cells in almost every conceivable way. Protein modifications can occur as a result of specific regulating actions of enzymes, such as tyrosine kinases phosphorylating tyrosine residues or by nonenzy...

Descripción completa

Detalles Bibliográficos
Autores principales: Heininen, Juho, Erbacher, Catharina, Kotiaho, Tapio, Kostiainen, Risto, Teppo, Jaakko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2023
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10243104/
https://www.ncbi.nlm.nih.gov/pubmed/37146082
http://dx.doi.org/10.1021/acs.jproteome.3c00061
_version_ 1785054359587913728
author Heininen, Juho
Erbacher, Catharina
Kotiaho, Tapio
Kostiainen, Risto
Teppo, Jaakko
author_facet Heininen, Juho
Erbacher, Catharina
Kotiaho, Tapio
Kostiainen, Risto
Teppo, Jaakko
author_sort Heininen, Juho
collection PubMed
description [Image: see text] Post-translational modifications (PTMs) alter the function and fate of proteins and cells in almost every conceivable way. Protein modifications can occur as a result of specific regulating actions of enzymes, such as tyrosine kinases phosphorylating tyrosine residues or by nonenzymatic reactions, such as oxidation related to oxidative stress and diseases. While many studies have addressed the multisite, dynamic, and network-like properties of PTMs, only little is known of the interplay of the same site modifications. In this work, we studied the enzymatic phosphorylation of oxidized tyrosine (l-DOPA) residues using synthetic insulin receptor peptides, in which tyrosine residues were replaced with l-DOPA. The phosphorylated peptides were identified by liquid chromatography-high-resolution mass spectrometry and the site of phosphorylation by tandem mass spectrometry. The results clearly show that the oxidized tyrosine residues are phosphorylated, displaying a specific immonium ion peak in the MS(2) spectra. Furthermore, we detected this modification in our reanalysis (MassIVE ID: MSV000090106) of published bottom-up phosphoproteomics data. The modification, where both oxidation and phosphorylation take place at the same amino acid, has not yet been published in PTM databases. Our data indicate that there can be multiple PTMs that do not exclude each other at the same modification site.
format Online
Article
Text
id pubmed-10243104
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher American Chemical Society
record_format MEDLINE/PubMed
spelling pubmed-102431042023-06-07 Enzymatic Phosphorylation of Oxidized Tyrosine Residues Heininen, Juho Erbacher, Catharina Kotiaho, Tapio Kostiainen, Risto Teppo, Jaakko J Proteome Res [Image: see text] Post-translational modifications (PTMs) alter the function and fate of proteins and cells in almost every conceivable way. Protein modifications can occur as a result of specific regulating actions of enzymes, such as tyrosine kinases phosphorylating tyrosine residues or by nonenzymatic reactions, such as oxidation related to oxidative stress and diseases. While many studies have addressed the multisite, dynamic, and network-like properties of PTMs, only little is known of the interplay of the same site modifications. In this work, we studied the enzymatic phosphorylation of oxidized tyrosine (l-DOPA) residues using synthetic insulin receptor peptides, in which tyrosine residues were replaced with l-DOPA. The phosphorylated peptides were identified by liquid chromatography-high-resolution mass spectrometry and the site of phosphorylation by tandem mass spectrometry. The results clearly show that the oxidized tyrosine residues are phosphorylated, displaying a specific immonium ion peak in the MS(2) spectra. Furthermore, we detected this modification in our reanalysis (MassIVE ID: MSV000090106) of published bottom-up phosphoproteomics data. The modification, where both oxidation and phosphorylation take place at the same amino acid, has not yet been published in PTM databases. Our data indicate that there can be multiple PTMs that do not exclude each other at the same modification site. American Chemical Society 2023-05-05 /pmc/articles/PMC10243104/ /pubmed/37146082 http://dx.doi.org/10.1021/acs.jproteome.3c00061 Text en © 2023 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Heininen, Juho
Erbacher, Catharina
Kotiaho, Tapio
Kostiainen, Risto
Teppo, Jaakko
Enzymatic Phosphorylation of Oxidized Tyrosine Residues
title Enzymatic Phosphorylation of Oxidized Tyrosine Residues
title_full Enzymatic Phosphorylation of Oxidized Tyrosine Residues
title_fullStr Enzymatic Phosphorylation of Oxidized Tyrosine Residues
title_full_unstemmed Enzymatic Phosphorylation of Oxidized Tyrosine Residues
title_short Enzymatic Phosphorylation of Oxidized Tyrosine Residues
title_sort enzymatic phosphorylation of oxidized tyrosine residues
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10243104/
https://www.ncbi.nlm.nih.gov/pubmed/37146082
http://dx.doi.org/10.1021/acs.jproteome.3c00061
work_keys_str_mv AT heininenjuho enzymaticphosphorylationofoxidizedtyrosineresidues
AT erbachercatharina enzymaticphosphorylationofoxidizedtyrosineresidues
AT kotiahotapio enzymaticphosphorylationofoxidizedtyrosineresidues
AT kostiainenristo enzymaticphosphorylationofoxidizedtyrosineresidues
AT teppojaakko enzymaticphosphorylationofoxidizedtyrosineresidues